Correction for Bos et al., Phytophthora infestans effector AVR3a is essential for virulence and manipulates plant immunity by stabilizing host E3 ligase CMPG1

PLANT BIOLOGY Correction for “Phytophthora infestans effector AVR3a is essential for virulence and manipulates plant immunity by stabilizing host E3 ligase CMPG1,” by Jorunn I. B. Bos, Miles R. Armstrong, Eleanor M. Gilroy, Petra C. Boevink, Ingo Hein, Rosalind M. Taylor, Tian Zhendong, Stefan Engelhardt, Ramesh R. Vetukuri, Brian Harrower, Christina Dixelius, Glenn Bryan, Ari Sadanandom, Stephen C. Whisson, Sophien Kamoun, and Paul R. J. Birch, which was first published May 10, 2010; 10.1073/pnas.0914408107 (Proc. Natl. Acad. Sci. U.S.A. 107, 9909–9914).

The authors note, “We mistakenly used the wrong lanes in Western blots shown in the original Fig. 1B. The objective of Fig 1B was to demonstrate that CMPG1 was stable when expressed in yeast. Critically, as no interaction was observed between CMPG1 and the AVR3aKI 60-146 (AVR3a^{KI Y147del}) mutant in Fig. 1A, the question arose: were CMPG1 and the effector variant stable? The corrected Fig. 1B demonstrates that this is indeed the case, and thus does not alter our original findings that a failure to interact is not due to protein instability in yeast. Additionally, in Fig. 4 we erroneously highlighted the wrong sides of leaves in Fig. 4 D–G. In the legend for this figure we should have indicated that the enhanced colonization of P.cinfestans silenced line CS12, caused by transient expression of pGRAB::Avr3a^EM or pGRAB::Avr3a^KI, is observed on the right side of each leaf. We apologize for these mistakes.

Fig. 1.

AVR3a^KI and AVR3a^EM interact with and stabilize CMPG1. (A) AVR3a^KI and AVR3a^EM with (21-147) and without (60-147; shown only for AVR3a^KI) RXLR-encoding portions interact with CMPG1 in Y2H (LacZ and -His reporter genes activated). Whereas AVR3a^KI/Y147del (Y147 deletion) and AVR3a^KI/Y147S mutants fail to interact with CMPG1, the AVR3a^KI/Y147F mutant interacts. (B) Western blots probed with anti-myc and anti-HA antibodies following expression of myc-CMPG1 in binding domain vector, with HA-AVR3a^EM (21-147), HA-AVR3a^KI (21-147), HA-AVR3a^KI/Y147del (60-146) activation domain (AD) fusions, or AD alone, in yeast (each indicated with a white star). Full-length immunoblots are shown in Fig. S8. (C) Western blots probed with anti-myc and anti-FLAG antibodies, showing a time-course of transient coexpression (by agroinfiltration) of FLAG-AVR3a^KI, FLAG-AVR3a^EM, FLAG- AVR3a^KI/Y147del or a vector control with 4x-myc-ΔN-StCMPG1a at 2 and 3 d postinoculation (dpi). (D) As in C, but at 5 dpi. (E) As in C but at 5 dpi with 4x-myc-StCMPG1b (full-length) and 4x-myc-ΔN-StCMPG1b (lacking the N-terminal 29 amino acids). Protein sizes are indicated in kDa. Protein loading is shown by Coomassie blue (CBB) or Ponceau S (PS) staining. (F) Fluorimeter measurements (in relative fluorescence units) following coexpression in N. benthamiana of the split YFP constructs CMPG1::N-YFP (YN), CMPG1::C-YFP (YC), C-YFP::AVR3a^KI, C-YFP::AVR3a^EM, C-YFP::AVR3a^KI/Y147del as indicated. (G) Confocal microscopy following coexpression in N. benthamiana of split YFP constructs CMPG1-YC with a vector expressing free N-YFP, and CMPG1-YC with N-YFP::AVR3a^KI, N-YFP::AVR3a^EM, and N-YFP::AVR3a^KI/Y147del constructs as indicated in the panels. (Scale bars, 200 μm.)

Fig. 4.

Avr3a is essential for virulence. (A) Infection of potato cultivar Bintje by wild-type P. infestans isolate 88069 (left leaf) and Avr3a-silenced line CS12 (right leaf) at 4 dpi. (B) Infection of N. benthamiana by wild-type P. infestans isolate 88069 at 6 dpi. (C) Infection of N. benthamiana by Avr3a-silenced line CS12 at 6 dpi. (D) Infection of N. benthamiana by Avr3a-silenced line CS12 at 6 dpi, following agroexpression of pGRAB::Avr3a^EM (right half of leaf) or empty pGRAB (left half of leaf). (E) As in D, but leaf is stained with trypan blue. (F) Infection of N. benthamiana by Avr3a-silenced line CS12 at 6 dpi, following agroexpression of pGRAB::Avr3a^KI (right half of leaf) or empty pGRAB (left half of leaf). (G) As in F, but stained with trypan blue. (H) Measurements at 6 dpi of lesion sizes (mm) following infection of isolate 88069 or silenced line CS12 on leaves that were uninfiltrated, infiltrated with agromix, agro-infiltrated to express AVR4a^KI, Avr3a^EM, or empty pGRAB vector. These complementation experiments were repeated on four occasions, with leaves from six plants for each treatment/construct.

The corrected Fig. 1B appears below, along with the corrected Fig. 4 legend, and the full-length immunoblots used in Fig. 1B are shown below as Fig. S8.

Fig. S8.

Full length immunoblots used for the revised Fig. 1B showing proteins revealed by anti-myc or anti-HA antibodies as indicated, with corresponding Ponceau stains (PS) shown below each immunoblot.